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High content imaging of relative ATP levels for mitochondrial toxicity prediction in human induced pluripotent stem cell derived cardiomyocytes

  • 0Merck & Co., Inc., Rahway, NJ 07065, USA.
Toxicology +

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February 19, 2025

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February 19, 2025

View abstract on PubMed

Summary

This summary is machine-generated.

We developed a new assay using human induced pluripotent stem cell derived cardiomyocytes (hiPSC-CMs) to detect mitochondrial toxicity from drugs by measuring cellular ATP levels. This method aids in identifying cardiotoxic compounds early.

Area Of Science

  • Cardiovascular Research
  • Stem Cell Biology
  • Toxicology

Background

  • Drug-induced cardiotoxicity is a major concern in therapeutic development.
  • Mitochondrial dysfunction is a key mechanism in some cardiotoxic effects.
  • Human induced pluripotent stem cell derived cardiomyocytes (hiPSC-CMs) are valuable models for cardiotoxicity screening.

Purpose Of The Study

  • To develop and validate a high-content imaging assay for assessing mitochondrial toxicity in hiPSC-CMs.
  • To investigate the utility of ATP-Red fluorescent dye for measuring subcellular ATP levels in response to toxicants.
  • To evaluate the assay's ability to distinguish between cardiotoxic and non-cardiotoxic compounds.

Main Methods

  • Utilized hiPSC-CMs and a fluorescent dye (ATP-Red) to monitor subcellular ATP levels in living cells.
  • Treated cells with known mitochondrial toxicants (antimycin, oligomycin) and clinically relevant drugs.
  • Assessed ATP levels over time and concentration, with and without glucose supplementation.
  • Validated ATP measurements against orthogonal methods like whole-cell ATP assays and mitochondrial membrane potential.

Main Results

  • Demonstrated time- and concentration-dependent decreases in ATP-Red signal with antimycin and oligomycin.
  • Showed that decreased ATP levels could be rescued by glucose supplementation, indicating reliance on glycolysis.
  • Identified decreased ATP levels in hiPSC-CMs treated with known mitochondrial toxicants but not with non-toxic compounds.
  • ATP measurements correlated well with other toxicity assays, with amiodarone as a notable exception in mitochondrial membrane potential readings.

Conclusions

  • The developed high-throughput imaging assay effectively assesses mitochondrial ATP dynamics in hiPSC-CMs.
  • This assay provides mechanistic insights into drug-induced cardiotoxicity by evaluating mitochondrial function.
  • The assay is a valuable tool for early-stage screening of potential cardiotoxic liabilities in new drug candidates.

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